Schwuchow J M, Kern V D, Wagner T, Sack F D
Department of Plant Biology, Ohio State University, Columbus, USA.
Protoplasma. 2000;211(3-4):225-33. doi: 10.1007/BF01304490.
Determinations of plant or algal cell density (cell mass divided by volume) have rarely accounted for the extracellular matrix or shrinkage during isolation. Three techniques were used to indirectly estimate the density of intact apical cells from protonemata of the moss Ceratodon purpureus. First, the volume fraction of each cell component was determined by stereology, and published values for component density were used to extrapolate to the entire cell. Second, protonemal tips were immersed in bovine serum albumin solutions of different densities, and then the equilibrium density was corrected for the mass of the cell wall. Third, apical cell protoplasts were centrifuged in low-osmolarity gradients, and values were corrected for shrinkage during protoplast isolation. Values from centrifugation (1.004 to 1.015 g/cm3) were considerably lower than from other methods (1.046 to 1.085 g/cm3). This work appears to provide the first corrected estimates of the density of any plant cell. It also documents a method for the isolation of protoplasts specifically from apical cells of protonemal filaments.
对植物或藻类细胞密度(细胞质量除以体积)的测定很少考虑到分离过程中的细胞外基质或收缩情况。我们使用了三种技术来间接估计来自紫萼藓原丝体顶端完整细胞的密度。首先,通过体视学确定每个细胞组分的体积分数,并使用已发表的组分密度值外推至整个细胞。其次,将原丝体顶端浸入不同密度的牛血清白蛋白溶液中,然后针对细胞壁的质量校正平衡密度。第三,将顶端细胞原生质体在低渗透压梯度中离心,并针对原生质体分离过程中的收缩对数值进行校正。离心法得到的值(1.004至1.015克/立方厘米)明显低于其他方法(1.046至1.085克/立方厘米)。这项工作似乎首次提供了对任何植物细胞密度的校正估计值。它还记录了一种专门从原丝体细丝顶端细胞分离原生质体的方法。